MATERIALS AND METHODS

Plant was collected from the forest regions of Thalakona of Chittoor district and was
authenticated by Mr. A. Laxma Reddy, Retired Professor, Dept. of Botany, Nagarjuna
Government College, Nalgonda. The plant herbarium was prepared and deposited in the
Department of Pharmacognosy, Nalanda College of Pharmacy for future reference. The plant
was identified as Limnophila indica (Linn.) Druce (Scrophulariaceae)
Procurement of Micro organisms:
Shigella flexneri MTCC 1457 was procured as freeze dried form from Microbial type culture
collection, Chandighar, India. Shigella dysentery and Shigella boydii were the two pathogenic
antibiotic resistant bacterial strains that were procured from Microbiology laboratory of
Kamineni Institute of Medical Sciences, Nalgonda, Andhra Pradesh, India.
Procurement of animals
Wistar Albino Rats of either sex weighing between 150-200g were procured from National
Institute of Nutrition, Hyderabad, A.P, India. The experimental protocol was approved from the
Institutes animal ethics committee under the reference no. NCOP/IAEC/approval/36/2011 and
then experimental studies were undergone according to their rules and regulations. The animals
were housed in metabolic cages, bedded with rice husk under standard environmental conditions
and had free access to standard pellet diet and water ad libitum.
Preparation of extract
The 500g of powered plant material was defatted with petroleum ether and then extracted with
methanol for 72 h at 45C. The extract thus obtained was then concentrated under vacuum using
rotary vacuum evaporator and then subjected to preliminary chemical screening to identify the
active chemical constituents.
Formulation of Herbal Effervescent Granules
Herbal Effervescent granules were prepared by wet granulation method. 11.78g citric acid,
23.56g tartaric acid, 40.05g sodium bicarbonate and 24.6g were triturated into fine powder. Then
sufficient alcohol was added to make a damp mass. This mass was passed through sieve no 10 to
get granules and these granules were dried in hot air oven at 40C and then they were packed in
air tight container.

Bahan dan Metode

Tanaman dikumpulkan dari daerah hutan Thalakona Kabupaten Chittoor dan
dikonfirmasi oleh Mr A. Laxma Reddy, Pensiunan Profesor, Departemen Botani, Nagarjuna
Pemerintah College, Nalgonda. Herbarium tanaman disiapkan dan disimpan di Departemen
Farmakognosi, Nalanda College of Pharmacy untuk referensi di masa mendatang. Tumbuhan ini
diidentifikasi sebagai Limnophila indica (Linn.) Druce (Scrophulariaceae)
Pengadaan organisme Mikro:
Shigella flexneri MTCC 1457 yang diperoleh sebagai bentuk kering beku dari koleksi kultur
mikroba jenis, Chandighar, India. Shigella disentri dan Shigella boydii adalah dua patogen
antibiotik strain bakteri resisten yang dibeli dari laboratorium Mikrobiologi Kamineni Institute of
Medical Sciences, Nalgonda, Andhra Pradesh, India.
Pengadaan hewan
Wistar Tikus Albino dari kedua jenis kelamin dengan berat antara 150-200g tersebut dibeli dari
National Institute of Nutrition, Hyderabad, AP, India. Protokol eksperimental telah disetujui dari
komite etika hewan Institutes bawah referensi no. NCOP/IAEC/approval/36/2011 dan kemudian
studi eksperimental yang mengalami sesuai dengan aturan dan peraturan. Hewan-hewan yang
tinggal di kandang metabolik, bersetubuh dengan sekam padi dalam kondisi lingkungan standar
dan memiliki akses gratis untuk diet pelet standar dan ad libitum.
Persiapan ekstrak
The 500g bahan tanaman powered dihilangkan lemaknya dengan petroleum eter dan kemudian
diekstraksi dengan metanol selama 72 jam pada 45 C. Ekstrak yang diperoleh kemudian
dipekatkan dalam vakum menggunakan rotary vacuum evaporator dan kemudian dilakukan
pemeriksaan kimia pendahuluan untuk mengidentifikasi kandungan kimia aktif.
Perumusan Butiran Effervescent Herbal
Butiran Effervescent Herbal dibuat dengan metode granulasi basah. 11.78g asam sitrat, asam
tartarat 23.56g, natrium bikarbonat 40.05g dan 24.6g yang ditriturasi menjadi bubuk halus.
Kemudian alkohol yang cukup ditambahkan untuk membuat massa lembab. Massa ini disahkan
melalui saringan no 10 untuk mendapatkan butiran dan butiran tersebut dikeringkan dalam oven
udara panas pada suhu 40 C dan kemudian mereka yang dikemas dalam wadah kedap udara.